Bioengineered Ferritin Nanocarriers for Cancer Therapy.

Ferritin naturally exists in most organisms and can specifically recognize the transferrin 1 receptor (TfR1), which is generally highly expressed on various types of tumor cells. The pH dependent reversible assembling and disassembling property of ferritin renders it as a suitable candidate for encapsulating a variety of anticancer drugs and imaging probes. Ferritins external surface is chemically and genetically modifiable which can serve as attachment site for tumor specific targeting peptides or moieties. Moreover, the biological origin of these protein cages makes it a biocompatible nanocarrier that stabilizes and protects the enclosed particles from the external environment without provoking any toxic or immunogenic responses. Recent studies, further establish ferritin as a multifunctional nanocarrier for targeted cancer chemotherapy and phototherapy. In this review, we introduce the favorable characteristics of ferritin drug carriers, the specific targeted surface modification and a multifunctional nanocarriers combined chemotherapy with phototherapy for tumor treatment. Taken together, ferritin is a potential ideal base of engineered nanoparticles for tumor therapy and still needs to explore more on its way.

Bio-Engineered Nisin with Increased Anti-Staphylococcus and Selectively Reduced Anti-Lactococcus Activity for Treatment of Bovine Mastitis.

Bovine mastitis is a significant economic burden for dairy enterprises, responsible for premature culling, prophylactic and therapeutic antibiotic use, reduced milk production and the withholding (and thus wastage) of milk. There is a desire to identify novel antimicrobials that are expressly directed to veterinary applications, do not require a lengthy milk withholding period and that will not have a negative impact on the growth of lactic acid bacteria involved in downstream dairy fermentations. Nisin is the prototypical lantibiotic, a family of highly modified antimicrobial peptides that exhibit potent antimicrobial activity against many Gram-positive microbes, including human and animal pathogens including species of Staphylococcus and Streptococcus. Although not yet utilized in the area of human medicine, nisin is currently applied as the active agent in products designed to prevent bovine mastitis. Over the last decade, we have harnessed bioengineering strategies to boost the specific activity and target spectrum of nisin against several problematic microorganisms. Here, we screen a large bank of engineered nisin derivatives to identify novel derivatives that exhibit improved specific activity against a selection of staphylococci, including mastitis-associated strains, but have unchanged or reduced activity against dairy lactococci. Three such peptides were identified; nisin A M17Q, nisin A T2L and nisin A HTK.

Bioengineering tools to speed up the discovery and preclinical testing of vaccines for SARS-CoV-2 and therapeutic agents for COVID-19.

This review provides an update for the international research community on the cell modeling tools that could accelerate the understanding of SARS-CoV-2 infection mechanisms and could thus speed up the development of vaccines and therapeutic agents against COVID-19. Many bioengineering groups are actively developing frontier tools that are capable of providing realistic three-dimensional (3D) models for biological research, including cell culture scaffolds, microfluidic chambers for the culture of tissue equivalents and organoids, and implantable windows for intravital imaging. Here, we review the most innovative study models based on these bioengineering tools in the context of virology and vaccinology. To make it easier for scientists working on SARS-CoV-2 to identify and apply specific tools, we discuss how they could accelerate the discovery and preclinical development of antiviral drugs and vaccines, compared to conventional models.

Common principles and best practices for engineering microbiomes.

Despite broad scientific interest in harnessing the power of Earth's microbiomes, knowledge gaps hinder their efficient use for addressing urgent societal and environmental challenges. We argue that structuring research and technology developments around a design-build-test-learn (DBTL) cycle will advance microbiome engineering and spur new discoveries of the basic scientific principles governing microbiome function. In this Review, we present key elements of an iterative DBTL cycle for microbiome engineering, focusing on generalizable approaches, including top-down and bottom-up design processes, synthetic and self-assembled construction methods, and emerging tools to analyse microbiome function. These approaches can be used to harness microbiomes for broad applications related to medicine, agriculture, energy and the environment. We also discuss key challenges and opportunities of each approach and synthesize them into best practice guidelines for engineering microbiomes. We anticipate that adoption of a DBTL framework will rapidly advance microbiome-based biotechnologies aimed at improving human and animal health, agriculture and enabling the bioeconomy.

Bioengineering of Piper longum L. extract mediated silver nanoparticles and their potential biomedical applications.

The present investigation highlights the strong antioxidant, anticancer and larvicidal potential of green synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Piper longum L. for their diverse therapeutic applications. The optimum conditions for the synthesis of AgNPs were recorded as 1 mM AgNO3, 60 ±â€¯2 °C at pH 6 for 120 min. Synthesized AgNPs proved to be highly stable and monodispersed as characterized through various techniques. UV-Vis spectrum of biosynthesized AgNPs showed a maximum absorption peak at 420 nm. Field emission-Scanning electron microscopy (FE-SEM) and High resolution-Transmission electron microscopy (HR-TEM) micrographs showed the spherical shape of AgNPs with mean diameter size of 28.8 nm. Existence of crystallographic AgNPs was proved by X-ray diffraction (XRD) pattern analysis. Presence of phenolics, terpenoids and flavonoids compounds which act as bioreducing agents were confirmed by Fourier-transform infrared spectroscopy (FTIR) analysis. Furthermore, the AgNPs and leaf extracts prepared individually in different solvents such as methanol, ethyl acetate, chloroform, hexane and aqueous were assessed for their bio-efficacies. AgNPs showed the enhanced antioxidant (IC50 67.56 µg) and radical-scavenging activities (IC50 196.8 µg) as compared to the crude leaf extracts. Anticancer activity revealed the strong and dose-dependent cytotoxic effect of AgNPs against the HeLa cells showing maximum IC50 value being 5.27 µg/mL after 24 h and was also found to be non-toxic to normal cells (HEK). The AgNPs induced the fragmentation of DNA in the cells, indicating the occurrence of apoptosis and necrosis. Subsequently, an efficient larvae mortality was also recorded against Anopheles stephensi having LC50 and LC90 values being 8.969 and 16.102 ppm, followed by Aedes aegypti (LC50;14.791 and LC90;28.526 ppm) and Culex quinquefasciatus (LC50;18.662 and LC90;40.903 ppm) after 72 h of exposure. Besides, they showed no toxicity against Mesocyclops thermocyclopoides (non-target organism). This is the first report showing strong anti-tumorous and larvicidal activity of AgNPs synthesized using P. longum leaf extract against cervical cancer cell line and mosquito vectors causing dengue, malaria and filariasis. Based on our findings, we suggest that AgNPs derived using P. longum leaf extract possessed excellent anti-cancerous and mosquito larvicidal potential and therefore, can be bioprospected further for the management of these hazardous health diseases. This study has given a new insight for the novel drug designing after conducting experiments on the in vivo models.

Biomimetic engineering endothelium-like coating on cardiovascular stent through heparin and nitric oxide-generating compound synergistic modification strategy.

Co-immobilization of two or more molecules with different and complementary functions to prevent thrombosis, suppress smooth muscle cell (SMC) proliferation, and support endothelial cell (EC) growth is generally considered to be promising for the re-endothelialization on cardiovascular stents. However, integration of molecules with distinct therapeutic effects does not necessarily result in synergistic physiological functions due to the lack of interactions among them, limiting their practical efficacy. Herein, we apply heparin and nitric oxide (NO), two key molecules of the physiological functions of endothelium, to develop an endothelium-mimetic coating. Such coating is achieved by sequential conjugation of heparin and the NO-generating compound selenocystamine (SeCA) on an amine-bearing film of plasma polymerized allylamine. The resulting surface combines the anti-coagulant (anti-FXa) function provided by the heparin and the anti-platelet activity of the catalytically produced NO. It also endows the stents with the ability to simultaneously up-regulate α-smooth muscle actin (α-SMA) expression and to increase cyclic guanylate monophosphate (cGMP) synthesis of SMC, thereby significantly promoting their contractile phenotype and suppressing their proliferation. Importantly, this endothelium-biomimetic coating creates a favorable microenvironment for EC over SMC. These features impressively improve the antithrombogenicity, re-endothelialization and anti-restenosis of vascular stents in vivo.

Bioengineering of microbial transglutaminase for biomedical applications.

Microbial transglutaminase (mTGase) is commonly known in the food industry as meat glue due to its incredible ability to "glue" meat proteins together. Aside from being widely exploited in the meat processing industries, mTGase is also widely applied in other food and textile industries by catalysing the formation of isopeptide bonds between peptides or protein substrates. The advancement of technology has opened up new avenues for mTGase in the field of biomedical engineering. Efforts have been made to study the structural properties of mTGase in order to gain an in-depth understanding of the structure-function relationship. This review highlights the developments in mTGase engineering together with its role in biomedical applications including biomaterial fabrication for tissue engineering and biotherapeutics.

Biomimetic cardiovascular platforms for in vitro disease modeling and therapeutic validation.

Bioengineered tissues have become increasingly more sophisticated owing to recent advancements in the fields of biomaterials, microfabrication, microfluidics, genetic engineering, and stem cell and developmental biology. In the coming years, the ability to engineer artificial constructs that accurately mimic the compositional, architectural, and functional properties of human tissues, will profoundly impact the therapeutic and diagnostic aspects of the healthcare industry. In this regard, bioengineered cardiac tissues are of particular importance due to the extremely limited ability of the myocardium to self-regenerate, as well as the remarkably high mortality associated with cardiovascular diseases worldwide. As novel microphysiological systems make the transition from bench to bedside, their implementation in high throughput drug screening, personalized diagnostics, disease modeling, and targeted therapy validation will bring forth a paradigm shift in the clinical management of cardiovascular diseases. Here, we will review the current state of the art in experimental in vitro platforms for next generation diagnostics and therapy validation. We will describe recent advancements in the development of smart biomaterials, biofabrication techniques, and stem cell engineering, aimed at recapitulating cardiovascular function at the tissue- and organ levels. In addition, integrative and multidisciplinary approaches to engineer biomimetic cardiovascular constructs with unprecedented human and clinical relevance will be discussed. We will comment on the implementation of these platforms in high throughput drug screening, in vitro disease modeling and therapy validation. Lastly, future perspectives will be provided on how these biomimetic platforms will aid in the transition towards patient centered diagnostics, and the development of personalized targeted therapeutics.

Antimicrobial Characterization of Advanced Materials for Bioengineering Applications.

The development of new advanced materials with enhanced properties is becoming more and more important in a wide range of bioengineering applications. Thus, many novel biomaterials are being designed to mimic specific environments required for biomedical applications such as tissue engineering and controlled drug delivery. The development of materials with improved properties for the immobilization of cells or enzymes is also a current research topic in bioprocess engineering. However, one of the most desirable properties of a material in these applications is the antimicrobial capacity to avoid any undesirable infections. For this, we present easy-to-follow protocols for the antimicrobial characterization of materials based on (i) the agar disk diffusion test (diffusion method) and (ii) the ISO 22196:2007 norm to measure the antimicrobial activity on material surfaces (contact method). This protocol must be performed using Gram-positive and Gram-negative bacteria and yeast to cover a broad range of microorganisms. As an example, 4 materials with different chemical natures are tested following this protocol against Staphylococcus aureus, Escherichia coli, and Candida albicans.The results of these tests exhibit non-antimicrobial activity for the first material and increasing antibacterial activity against Gram-positive and Gram-negative bacteria for the other 3 materials. However, none of the 4 materials are able to inhibit the growth of Candida albicans.

Synthetic Biology and Engineered Live Biotherapeutics: Toward Increasing System Complexity.

Recent advances in synthetic biology and biological system engineering have allowed the design and construction of engineered live biotherapeutics targeting a range of human clinical applications. In this review, we outline how systems approaches have been used to move from simple constitutive systems, where a single therapeutic molecule is expressed, to systems that incorporate sensing of the in vivo environment, feedback, computation, and biocontainment. We outline examples where each of these capabilities are achieved in different human disorders, including cancer, inflammation, and metabolic disease, in a number of environments, including the gastrointestinal tract, the liver, and the oral cavity. Throughout, we highlight the challenges of developing microbial therapeutics that are both sensitive and specific. Finally, we discuss how these systems are leading to the realization of engineered live biotherapeutics in the clinic.

Bioengineered silver nanoparticles as potent anti-corrosive inhibitor for mild steel in cooling towers.

Silver nanoparticle-aided enhancement in the anti-corrosion potential and stability of plant extract as ecologically benign alternative for microbially induced corrosion treatment is demonstrated. Bioengineered silver nanoparticles (AgNPs) surface functionalized with plant extract material (proteinacious) was generated in vitro in a test tube by treating ionic AgNO3 with the leaf extract of Azadirachta indica that acted as dual reducing as well as stabilizing agent. Purity and crystallinity of the AgNPs, along with physical and surface characterizations, were evaluated by performing transmission electron microscopy, Fourier transform infrared spectroscopy, energy dispersive x-ray spectra, single-area electron diffractions, zeta potential, and dynamic light scattering measurements. Anti-corrosion studies against mild steel (MS1010) by corrosion-inducive bacterium, Bacillus thuringiensis EN2 isolated from cooling towers, were evaluated by performing electrochemical impedance spectroscopy (EIS), weight loss analysis, and surface analysis by infrared spectroscopy. Our studies revealed that AgNPs profoundly inhibited the biofilm on MS1010 surface and reduced the corrosion rates with the CR of 0.5 mm/y and an inhibition efficiency of 77% when compared to plant extract alone with a CR of 2.2 mm/y and an inhibition efficiency of 52%. Further surface analysis by infrared spectra revealed that AgNPs formed a protective layer of self-assembled film on the surface of MS1010. Additionally, EIS and surface analysis revealed that the AgNPs have inhibited the bacterial biofilm and reduced the pit on MS1010. This is the first report disclosing the application of bioengineered AgNP formulations as potent anti-corrosive inhibitor upon forming a protective layer over mild steel in cooling water towers. Graphical Abstract ᅟ.

A co-culture microtunnel technique demonstrating a significant contribution of unmyelinated Schwann cells to the acceleration of axonal conduction in Schwann cell-regulated peripheral nerve development.

Schwann cells (SCs) contribute to the regulation of axonal conduction in a myelin-dependent and -independent manner. However, due to the lack of investigative techniques that are able to record axonal conduction under conditions that control the proliferation of specific SC types, little is known about the extent to which myelinated SCs (mSCs) and unmyelinated SCs (umSCs) modulate axonal conduction. In this study, a microtunnel-electrode approach was applied to a neuron/SC co-culture technique. Rat dorsal root ganglion neurons and SCs were co-cultured in a microtunnel-electrode device, which enabled recording of the conduction delay in multiple axons passing through microtunnels. Despite the absence of nuclei in the microtunnel when SCs were eliminated, cultured cells were densely packed and expressed S100 beta (an SC marker) at a rate of 96% in neuron/SC co-culture, indicating that SCs migrated into the microtunnel. In addition, supplementation with ascorbic acid after 6 days in vitro (DIV) successfully induced myelination from 22 DIV. Activity recording experiments indicated that the conduction delay decreased with culture length from 17 DIV in the neuron/SC co-culture but not in neuron monoculture. Interestingly, the SC-modulated shortening of conduction delay was attenuated at 17 DIV and 22 DIV by supplementing the culture medium with ascorbic acid and, at the same time, suppressing SC proliferation, suggesting that immature umSCs increased axonal conduction velocity in a cell density-dependent manner before the onset of myelination. These results suggest that this method is an effective tool for investigating the contributions of mSCs or umSCs to the regulation of axonal conduction.

Decreasing but still significant facilitation effect of cold-season macrophytes on wetlands purification function during cold winter.

To identify the facilitation effect of a cool-season aquatic macrophyte (FEam) for use in effluent purification via constructed floating wetlands (CFWs) and to determine the possible pathways used during a winter period with an average temperature of less than 5 °C, pilot-scale CFWs were planted with the cold-season macrophyte Oenanthe clecumbens and were operated as batch systems. Although some leaves withered, the roots retained relatively high levels of activity during the winter, which had average air and water temperatures of 3.63 and 5.04 °C, respectively. The N and P removal efficiencies in CFWs decreased significantly in winter relative to those in late autumn. The presence of cool-season plants resulted in significant improvements in N and P removal, with a FEam of 15.23-25.86% in winter. Microbial N removal accounted for 71.57% of the total N removed in winter, and the decrease in plant uptake was the dominant factor in the wintertime decrease in N removal relative to that in late autumn. These results demonstrate the importance of cold-season plants in CFWs for the treatment of secondary effluent during cold winters.

A novel process for obtaining pinosylvin using combinatorial bioengineering in Escherichia coli.

Pinosylvin as a bioactive stilbene is of great interest for food supplements and pharmaceuticals development. In comparison to conventional extraction of pinosylvin from plant sources, biosynthesis engineering of microbial cell factories is a sustainable and flexible alternative method. Current synthetic strategies often require expensive phenylpropanoic precursor and inducer, which are not available for large-scale fermentation process. In this study, three bioengineering strategies were described to the development of a simple and economical process for pinosylvin biosynthesis in Escherichia coli. Firstly, we evaluated different construct environments to give a highly efficient constitutive system for enzymes of pinosylvin pathway expression: 4-coumarate: coenzyme A ligase (4CL) and stilbene synthase (STS). Secondly, malonyl coenzyme A (malonyl-CoA) is a key precursor of pinosylvin bioproduction and at low level in E. coli cell. Thus clustered regularly interspaced short palindromic repeats interference (CRISPRi) was explored to inactivate malonyl-CoA consumption pathway to increase its availability. The resulting pinosylvin content in engineered E. coli was obtained a 1.9-fold increase depending on the repression of fabD (encoding malonyl-CoA-ACP transacylase) gene. Eventually, a phenylalanine over-producing E. coli consisting phenylalanine ammonia lyase was introduced to produce the precursor of pinosylvin, trans-cinnamic acid, the crude extraction of cultural medium was used as supplementation for pinosylvin bioproduction. Using these combinatorial processes, 47.49 mg/L pinosylvin was produced from glycerol.

Artificial Organs 2015: A Year in Review.

In this Editor's Review, articles published in 2015 are organized by category and briefly summarized. We aim to provide a brief reflection of the currently available worldwide knowledge that is intended to advance and better human life while providing insight for continued application of technologies and methods of organ Replacement, Recovery, and Regeneration. As the official journal of The International Federation for Artificial Organs, The International Faculty for Artificial Organs, the International Society for Rotary Blood Pumps, the International Society for Pediatric Mechanical Cardiopulmonary Support, and the Vienna International Workshop on Functional Electrical Stimulation, Artificial Organs continues in the original mission of its founders "to foster communications in the field of artificial organs on an international level." Artificial Organs continues to publish developments and clinical applications of artificial organ technologies in this broad and expanding field of organ Replacement, Recovery, and Regeneration from all over the world. We take this time also to express our gratitude to our authors for providing their work to this journal. We offer our very special thanks to our reviewers who give so generously of their time and expertise to review, critique, and especially provide meaningful suggestions to the author's work whether eventually accepted or rejected. Without these excellent and dedicated reviewers, the quality expected from such a journal could not be possible. We also express our special thanks to our Publisher, John Wiley & Sons for their expert attention and support in the production and marketing of Artificial Organs. We look forward to reporting further advances in the coming years.

Helicobacter pylori infection: A brief overview on alternative natural treatments to conventional therapy.

Helicobacter pylori is a human gastric pathogen considered as the etiologic agent of several gastric disorders, that may range from chronic gastritis to more severe outcomes, including gastric cancer. The current therapeutic scheme relies on the combination of several pharmacological substances, namely antibiotics and proton pump inhibitors. However, the cure rates obtained have been declining over the years, mostly due to bacterial resistance to antibiotics. In this context, the use of non-antibiotic substances is of the utmost importance regarding H. pylori eradication. In this review, we present different classes of compounds obtained from natural sources that have shown to present anti-H. pylori potential; we briefly highlight their possible use in the context of developing new therapeutic approaches.

Human induced pluripotent stem cell-derived cardiomyocytes: insights into molecular, cellular, and functional phenotypes.

Disease models are essential for understanding cardiovascular disease pathogenesis and developing new therapeutics. The human induced pluripotent stem cell (iPSC) technology has generated significant enthusiasm for its potential application in basic and translational cardiac research. Patient-specific iPSC-derived cardiomyocytes offer an attractive experimental platform to model cardiovascular diseases, study the earliest stages of human development, accelerate predictive drug toxicology tests, and advance potential regenerative therapies. Harnessing the power of iPSC-derived cardiomyocytes could eliminate confounding species-specific and interpersonal variations and ultimately pave the way for the development of personalized medicine for cardiovascular diseases. However, the predictive power of iPSC-derived cardiomyocytes as a valuable model is contingent on comprehensive and rigorous molecular and functional characterization.

Duration and magnitude of myofascial release in 3-dimensional bioengineered tendons: effects on wound healing.

CONTEXT: Myofascial release (MFR) is one of the most commonly used manual manipulative treatments for patients with soft tissue injury. However, a paucity of basic science evidence has been published to support any particular mechanism that may contribute to reported clinical efficacies of MFR. OBJECTIVE: To investigate the effects of duration and magnitude of MFR strain on wound healing in bioengineered tendons (BETs) in vitro. METHODS: The BETs were cultured on a deformable matrix and then wounded with a steel cutting tip. Using vacuum pressure, they were then strained with a modeled MFR paradigm. The duration of MFR dose consisted of a slow-loading strain that stretched the BETs 6% beyond their resting length, held them for 0, 1, 2, 3, 4, or 5 minutes, and then slowly released them back to baseline. To assess the effects of MFR magnitude, the BETs were stretched to 0%, 3%, 6%, 9%, or 12% beyond resting length, held for 90 seconds, and then released back to baseline. Repeated measures of BET width and the wound's area, shape, and major and minor axes were quantified using microscopy over a 48-hour period. RESULTS: An 11% and 12% reduction in BET width were observed in groups with a 9% (0.961 mm; P<.01) and 12% (0.952 mm; P<.05) strain, respectively. Reduction of the minor axis of the wound was unrelated to changes in BET width. In the 3% strain group, a statistically significant decrease (-40%; P<.05) in wound size was observed at 24 hours compared with 48 hours in the nonstrain, 6% strain, and 9% strain groups. Longer duration of MFR resulted in rapid decreases in wound size, which were observed as early as 3 hours after strain. CONCLUSION: Wound healing is highly dependent on the duration and magnitude of MFR strain, with a lower magnitude and longer duration leading to the most improvement. The rapid change in wound area observed 3 hours after strain suggests that this phenomenon is likely a result of the modification of the existing matrix protein architecture. These data suggest that MFR's effect on the extracellular matrix can potentially promote wound healing.